Dataset Information

Comparative computer-assisted sperm analysis in non-human primates.

ABSTRACT:

Background

Biomedical research has recently focused on developing new models of human disease by implementing genome-editing strategies in non-human primates (NHPs) to introduce relevant gene mutations. There is a need to establish objective semen evaluation methods to select sires for in vitro fertilization to perform germline editing in embryos.

Methods

Sperm motility kinematic parameters were evaluated using a computer-assisted semen analysis (CASA) instrument for rhesus macaques (Macaca mulatta), cynomolgus macaques (Macaca fascicularis), and common marmosets (Callithrix jacchus).

Results

Normative sperm kinematic parameters were established, revealing differences between marmosets and macaques. The impact of season on rhesus macaque sperm motility was modest, where changes in sperm motility related to season were dependent on the individual male.

Conclusions

These data provide a baseline of normative kinematic parameters for three captive NHP species, in which implementation of CASA may serve as a tool to evaluate NHP semen quality.

SUBMITTER: Schmidt JK

PROVIDER: S-EPMC7969417 | biostudies-literature | 2021 Apr

REPOSITORIES: biostudies-literature

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Publications

Comparative computer-assisted sperm analysis in non-human primates.

Schmidt Jenna K JK Mean Katherine D KD Dusek Brittany M BM Hinkle Hayly M HM Puntney Riley C RC Alexander Eric S ES Malicki Kerri B KB Sneed Emily L EL Moy Amy W AW Golos Thaddeus G TG

Journal of medical primatology 20210120 2

<h4>Background</h4>Biomedical research has recently focused on developing new models of human disease by implementing genome-editing strategies in non-human primates (NHPs) to introduce relevant gene mutations. There is a need to establish objective semen evaluation methods to select sires for in vitro fertilization to perform germline editing in embryos.<h4>Methods</h4>Sperm motility kinematic parameters were evaluated using a computer-assisted semen analysis (CASA) instrument for rhesus macaqu ...[more]

PMID: 33469948

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Project description:Semen motility is the most widely recognized semen quality parameter used by Artificial Insemination (AI) centers. With the increasing worldwide export of semen between AI centers there is an increasing need for standardized motility assessment methods. Computer-Assisted Sperm Analysis (CASA) technology is thought to provide an objective motility evaluation; however, results can still vary between laboratories. The aim of present study was to verify the impact of different setting values of the CASA IVOS II on motility, concentration, and morphology of bovine semen samples frozen in an extender with or without egg yolk and then decide on optimal settings for a further validation step across AI centers. Semen straws from 30 different bulls were analyzed using IVOS II with twelve modified settings. No significant changes were observed in semen concentration, percentage of motile sperm or kinetic results for either extender type. However, increasing settings for both STR and VAP progressive (%) from Low, Medium, and High cut-off values significantly (p<0.05) reduced the percentage of detected progressive spermatozoa, in egg yolk extender from 49.5±15.2, 37.2±11.9 to 11.9±5.3%, and in clear extender from 51.9±9.1, 35.8±7.3 to 10.0±2.4%, respectively. In clear extender only, the modification of droplet proximal head length significantly affected the detection of normal sperm percentages (88.0± 4.7 to 95.0±0.6 and 96.0±0.6%) and of the percentage of detected proximal droplets (12.2±4.7, 2.5±2.7 to 0.6±0.2%) for Low, Medium and High values respectively (p<0.05). The identification of sensitivity within the CASA system to changes in set parameters then led to the determination of an optimal IVOS II setting. The existing variability among centers for these phenotypes was reduced when the standardized settings were applied across different CASA units. The results clearly show the importance of applied settings for the final CASA results and emphasize the need for standardized settings to obtain comparable data.

Project description:BackgroundSperm quality, including semen volume, sperm count, concentration, and total and progressive motility (collectively, "semen parameters"), has declined in the recent decades. Computer-assisted sperm analysis (CASA) provides sperm kinematic parameters, and the temporal trends of which remain unclear. Our objective is to examine the temporal trend of both semen parameters and kinematic parameters in Shanghai, China, in the recent years.MethodsThis retrospective study analyzed semen parameters and kinematic parameters of 49,819 men attending our reproductive center by using CASA during 2015-2021. The total sample was divided into two groups: samples that surpassed the WHO guideline (2010) low reference limits ("above reference limit" group, ARL; n = 24,575) and samples that did not ("below reference limit" group, BRL; n = 24,614). One-way analysis of variance, Kruskal-Wallis test, independent samples t-test, and covariance analysis were used to assess the differences among groups. Year, age, and abstinence time were included in the multiple linear regression model of the ARL group to adjust the confounders and depict the trends in sperm quality.ResultsAmong all the total sample and the ARL and BRL groups, the age of subjects increased in recent years. Semen volume and sperm count showed declined tendency with years in the total sample, the ARL and BRL groups, and the subgroup of age or abstinence time, whereas sperm velocities showed increased tendency with years on the contrary. The multiple linear regression model of the ARL group, adjusting for age and abstinence time, confirmed these trends. Semen volume (β1= -0.162; CI: -0.172, -0.152), sperm count (β1= -9.97; CI: -10.813, -9.128), sperm concentration (β1 = -0.535; CI: -0.772, -0.299), motility (β1 = -1.751; CI: -1.830, -1.672), and progressive motility (β1 = -1.12; CI: -0.201, -0.145) decreased with year, whereas curvilinear line velocity (VCL) (β1 = 3.058; CI: 2.912, 3.203), straight line velocity (VSL) (β1 = 2.075; CI: 1.990, 2.161), and average path velocity (VAP) (β1 = 2.305; CI: 2.224, 2.386) increased over time (all p < 0.001). In addition, VCL, VSL, and VAP significantly declined with age and abstinence time.ConclusionThe semen parameters declined, whereas the kinematic parameters increased over the recent years. We propose that, although sperm count and motility declined over time, sperm motion velocity increased, suggesting a possible compensatory mechanism of male fertility.

Project description:The reduction of spermatozoa survival time is a major problem of canine chilled sperm for artificial insemination. The aim of the study was to improve the quality of canine chilled sperm during storage time. We therefore, evaluated the effects of eight treatments with different levels of soybean lecithin concentration (1, 3 and 5%) and egg yolk (20%) in Tris-citric-fructose or Tris-citric-fructose-mineral salts extender on chilled canine sperm quality during 10 days of storage. The sperm motility was analysed by computer-assisted sperm analysis (CASA), whereas plasma membrane integrity, acrosome membrane integrity and mitochondrial membrane potential parameters were determined using a fluorescent staining combination of propidium iodide (PI), Hoechst 33342 (H342), fluorescein isothiocyanate-conjugated Pisum sativum agglutinin (FITC-PSA) and 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolyl-carbocyanine iodide (JC-1) by confocal laser scanning microscope. The results showed that egg yolk was found to be better than soybean lecithin in Tris-citric-fructose or Tris-citric-fructose-mineral salts extender for maintaining the quality of chilled canine sperm within 10 days of storage (P < 0.05). Although egg yolk in Tris-citric-fructose extender could maintain the motility better than other extenders, egg yolk in Tris-citric-fructose-mineral salts extender was the highest in intact plasma membrane, intact acrosome membrane and high mitochondrial membrane potential (P < 0.05). In contrast, the sperm quality of soybean lecithin in Tris-citric-fructose-mineral salts extender was lower than that of soybean lecithin in Tris-citric-fructose extender, and soybean lecithin 1% was greater than soybean lecithin 3% and 5% in plasma membrane integrity, acrosome membrane integrity and mitochondrial membrane potential (P < 0.05). In conclusion, soybean lecithin cannot replace egg yolk in Tris-citric-fructose or Tris-citric-fructose-mineral salts extenders, and egg yolk in Tris-citric-fructose-mineral salts extender is superior to other extenders in chilling canine sperm.

Project description:Background and aimsDue to the rapid motility of the selected sperm, sperm parameters cannot be accurately determined by the manual method. So, the application of a computer-assisted sperm analysis system with a high frame rate (HFR-CASA, 85 Hz) in sperm selection is investigated.MethodsA total of 177 semen samples were collected for sperm selection with density gradient centrifugation. Then, the manual method and HFR-CASA method will be used to observe and analyze the sperm concentration, motility, and percentage of progressively motile sperm (PR) of the selected sperm samples. The quality control of sperm concentration was performed with microballoons. Two selected sperm samples were analyzed 10 times repeatedly to evaluate the accuracy of the HFR-CASA.ResultsThe results of microballoons analyzed with the HFR-CASA were in control. The coefficients of variation of sperm concentration, motility, and PR from two selected sperm samples were all below 10%. The results of 177 selected sperm samples analyzed by the manual method and HFR-CASA showed that although there were significant positive correlations in sperm concentration, motility, and PR between them (p < 0.001), the manual method significantly underestimated sperm concentration (p = 0.002) but overestimated sperm motility and PR (p < 0.001). When sperm concentration was below 50 × 106/mL, the manual method significantly overestimated sperm concentration (p < 0.05). However, when sperm concentration was more than 100 × 106/mL, the manual method significantly underestimated sperm concentration (p < 0.001). Regardless of sperm concentration, the manual method consistently overestimated sperm motility and PR (p < 0.001). When sperm concentration was more than 20 × 106/mL, there was no correlation in sperm PR between them (p > 0.05). When sperm concentration was below 50 × 106/mL, the correct rate of captured sperm by the HFR-CASA was more than 98%.ConclusionThe HFR-CASA method is more accurate than the manual method in analyzing the selected sperm samples.

Dataset Information

Comparative computer-assisted sperm analysis in non-human primates.

Background

Methods

Results

Conclusions

Publications

Comparative computer-assisted sperm analysis in non-human primates.

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